Matrix comprising a bioactive component containing phospholipid

ABSTRACT

The present invention concerns a matrix with a bioactive component containing phospholipid which is composed of 5 to 98% by weight phosphatidyl serine and 1 to 90% by weight phosphatidyl choline. In addition 1 to 94% by weight of a fat component of vegetable and/or animal origin, a wax component, a polyalcohol component and/or other physiologically compatible additives can be contained. In this connection a matrix is preferred which has a permanently flexible water-containing coat and which has a total diameter of between 0.3 to 20 mm. This matrix is used especially to strengthen the ability to cope with physical and mental stress and functional capacity.

The present invention concerns a matrix with a bioactive componentcontaining phospholipid as well as the use thereof.

The substance class of phospholipids are so-called complex lipids havingamphiphilic properties i.e. they are at the same time lipophilic andhydrophilic which, among others, enables them to form lipid bilayers inaqueous media.

Phospholipids (also referred to as phosphatides) are chemicallyconsidered phosphodiesters in which the phosphoric acid is esterified,on the one hand, with a sphingosine or glyceride residue and, on theother hand, with choline, ethanolamine, serine, inositol or glycerol.Phosphatidyl choline is also known as lecithin and at the same time isan eponym for a large group of special phospholipids, the lecithins.Phosphatidyl serine and phosphatidyl ethanolamine are also referred toas cephalins.

The lyso derivatives which also belong to this group are formed byhydrolytic cleavage by means of specific phospholipases.

Phospholipid-containing capsules are sufficient well-known from theprior art and contain phospholipids mostly as a coating substance. Ifphospholipids are used in the filling, i.e. in the core of the capsule,they mostly act there in small amounts as a formulation adjuvant usuallyhaving solubilizing properties.

As a result of their amphiphilic properties phospholipids are also usedas coating substances of the known liposomes and transferosomes. In thisconnection they are used especially in the field of mucosal applicationsdue to their bioadhesive properties where they are introduced especiallyinto nasal and oral cavities.

However, in a chemically-modified form phospholipids are also used assurface-active formulation adjuvants (surfactants).

It is also known that vesicles which carry phospholipids as a coat canbe produced by means of ultrasound.

Special granulates with lecithin coats are known from the Japaneseapplication JP 91 47 043 as well as from EP-A 493 441. These granulateswhich, among others, contain steroids as bioactive substances are usedas feed additives.

According to WO 87/04347 lyso-phospholipids are described assolubilizers for hydrophobic bioactive substances.

Pharmaceutical forms that can enter the lungs which use organic halogencompounds as a carrier for the phosphatidyl choline are described in theInternational Applications WO 99/16419 and 99/16421.

Soft gelatin capsules which are commercially available as KAL®-Lecithinand which contain 1200 mg soybean lecithin contain lecithin as abioactive ingredient. However, in order to accommodate this amount oflecithin in a capsule, capsule sizes have to be selected which comeclose to the centimetre limit and thus give rise to a limitedcompliance.

A process for producing phosphatidyl serine (PS), i.e. a phospholipid,is known from the German Patent DE 199 17 249. In this connection it isstated that the PS or corresponding PS products obtained in this mannercan be stabilized in aqueous systems by embedding them in a hard fat.However, the proposals made there are limited to soft gelatine capsuleswhich should have the special PS in their contents. Although thedescribed system stabilizes phosphatidyl serine which is known to behydrolytically unstable, it has the disadvantage that this formulationcannot be encapsulated.

Formulations of phosphatidyl serine and phosphatidyl choline especiallyin a mixture with other lecithins and/or oils have proven to be notsufficiently stable in soft capsules. Also there are very tightconstraints on the mixing (blending) with regard to the melting pointand flow behaviour of the capsule contents due to the technicalrequirements of the encapsulation which is effected typically by meansof a so-called rotary dye. Also the processing temperatures in theencapsulation process often have an adverse effect on the properties ofthe capsule coat and/or the capsule contents.

One result of this described instability of the lecithins as capsulecontents is that at best moderately pasty blends can be encapsulatedinstead of a desired liquid formulation.

In addition the encapsulation of phospholipids in general causes majorproblems since, as described, they also act as emulsifiers and thusrapidly effect in a mixing of the not yet hardened (dried) coat with thecontents during the encapsulation process. As a result the capsulesbecome permeable within a relatively short time, they leak and can thusno longer be used.

Hence the object of the present invention was to provide a matrix with abioactive component containing phospholipid which does not have thedisadvantages of the described prior art and which can be formulated inan economically acceptable manner. Moreover, the bioactive phospholipidcomponent introduced into the matrix should have an adequate stabilityfor the most common purposes of application even in an encapsulatedstate.

This object was achieved by an appropriate matrix which contains as abioactive component 5 to 98% by weight phosphatidyl serine (PS) and 1 to90% by weight phosphatidyl choline (PC) and in addition 1 to 94% byweight of at least one other component from the series of fat componentof vegetable and/or animal origin, wax component, polyalcohol componentand other physiologically compatible additives.

Surprisingly with the matrix according to the invention it has turnedout that the phosphatidyl serine and/or phosphatidyl choline portionscontained therein are extremely stable towards the otherwise negativehydrolysis and/or the general degradation of encapsulated lecithins.This is especially strongly pronounced in blends that are highly viscousand additionally have thixotropic properties. Previously it was onlyknown that highly viscous blends make lecithins more stable but it wasthen no longer possible to encapsulate these highly viscous mixtures(cf. DE-OS 199 17 249). This is even more surprising since it is knownespecially that phosphatidyl serine is significantly less stable thanother phospholipids.

In addition to the head group (i.e. serine and/or choline), thecompounds preferably each contain a residue at positions sn-1 and/orsn-2 which is derived from a C₂-C₃₀ carboxylic acid, in particular aC₁₂-C₂₈ carboxylic acid bound to the hydroxyl groups of the glycerol.The acid residues can be linear or branched, saturated ormonounsaturated or polyunsaturated. Particularly preferred residues areresidues that are formed by the binding of acetic acid, butyric acid,caproic acid, caprylic acid, capric acid, lauric acid, myristic acid,arachidic acid, behenic acid, lignoceric acid, β-linolenic acid,eicosapentaenoic acid, erucic acid, nervonic acid, α- or β-eleostericacid or parinaric acid. Residues are particularly preferred which areformed by binding of palmitic acid, stearic acid, oleic acid, linoleicacid, α-linolenic acid, arachidonic acid or docosahexaenoic acid. Theacid residues bound to the OH groups of the glycerol that are stillavailable can thereby be identical or different.

In the total matrix according to the invention phosphatidyl serine andphosphatidyl choline represent the bioactive part; the portions of theother components give the total matrix its advantageous properties.

In the context of the present invention the term “bioactive” isunderstood as the effect of phosphatidyl serine and phosphatidyl cholineto display a biological action in the living organism which usuallyapplies to corresponding preparations in the human or veterinary fieldduring or after their release from the total matrix in the resorptionarea, on the transport path or at the site of action. In this connectionthis bioactive effect is of course not limited to the two saidphospholipids but can also be displayed by the other components involvedin the total matrix. However, their bioactive effect is not of primaryimportance for the matrix according to the invention.

In the present context a matrix has proven to be particularly suitablewhose bioactive component contains 10 to 40% by weight and particularlypreferably 15 to 30% by weight phosphatidyl serine. A matrix is alsoparticularly suitable whose bioactive component contains 2.0 to 20% byweight phosphatidyl choline.

Among the relatively broad spectrum of possible compositions, acomposition has proven to be particularly advantageous for the claimedmatrix which is composed of 10 to 70% by weight and particularlypreferably 20 to 50% by weight of the fat component and/or 3 to 30% byweight and particularly preferably 5 to 20% by weight of the waxcomponent, 1 to 30% by weight and particularly preferably 1 to 20% byweight of the polyalcohol component and/or 1.0 to 5% by weight of otherphysiologically compatible additives.

In addition to the bioactive components phosphatidyl serine andphosphatidyl choline that are essential according to the invention, thematrix can contain as the fat component preferably refined, hydrogenatedand/or fractionated fats and in particular those that are rich inomega-3 and/or omega-6 fatty acids such as docosahexaenoic acid,arachidonic acid, eicosapentaenoic acid and conjugated linolenic acid,free fatty acids, in particular omega-3 and omega-6 fatty acids, as thewax component preferably bee wax, candellila wax, shellac, paraffin,monoglycerides or diglycerides and as the polyalcohol componentpreferably polyethylene glycol, polysorbate, polyglycerol esters, sugaresters or sorbitan esters. The matrix can also advantageously containtocopherols and derivatives thereof, tocotrienols and derivativesthereof, polycosanols and derivatives thereof, vitamins such as vitaminC and E also in a derivatized form, amino acids in particular theessential, branched and non-proteinogenic amino acids such as theanine,amino acid derivatives such as creatine, taurine, carnitine,phytosterols and derivatives thereof, (poly)phenolic compounds andderivatives thereof such as catechol, phenolic acids such as gallicacid, hydroxycinnamic acids, coumarins, (iso)flavonoids such asquercetin or genistein, lignans and lignins as well as tannin, saponins,mono-, sesqui- and di-terpenes, carotinoids such as beta-carotin, luteinor lycopin, glucosinolates, roughage such as non-starch polysaccharides,extracts of vegetable and/or animal origin, physiologically activeproteins such as lactoferrin and glycomacropeptide, phospholipids andglycolipids such as sphingosine or (phyto)sphingomyelin and/or mineralcomponents but also other suitable bioactive components which is alsotaken into consideration by the present invention.

Phosphatidyl serine and phosphatidyl choline are in particularstabilized according to the invention by other matrix components whichare selected such that the total matrix (i.e. composed of PS/PC and theother components) is solid at room temperature and namely to such anextent that when using fats (triglycerides) the solid proportion of thetriglyceride that can be determined by TLC is >80% at 23° C. In additionthe components are advantageously selected such that the total matrixexhibits the property of shear dilution which for example can beachieved by the preferred use of a combination of fat and wax (e.g. beewax) in conjunction with PC/PS in the matrix when the triglyceridecontains a sufficiently high proportion of solid i.e. unmeltedtriglycerides. In this case preferred matrix components have aproportion of saturated fatty acids of more than 50% and advantageouslyno more than four mainly occurring triglyceride species are present. Inthis connection the use of palm kernel oil together with bee wax hasproven to be particularly advantageous.

Hence a person skilled in the art can readily select matrices that aresuitable according to the invention on the basis of the criteria 1) thesolid proportion of the triglyceride that can be determined by TLCis >80% at 23° C. and 2) the proportion of saturated fatty acids is morethan 50%.

In another embodiment of the invention the phosphatidyl serine andphosphatidyl choline are stabilized by using a matrix which contains apolyalcohol component. This matrix can be overall somewhat more liquidwhere in particular glycerol is added as a polyalcohol component.Disadvantages of conventional lecithin matrices can be overcome in thisembodiment. In lecithin matrices the water content is always low due tothe strong lipophilic properties of the matrix and the water that ispresent does not move freely in the matrix but is bound to the polarhead groups and hydrates them and/or hydrolyses the head group. Thisresults in the instability of the phospholipid (i.e. PC and/or PS) inconventional matrices. By the addition according to the invention of apolyalcohol component, for example glycerol as a polar substance thewater from the head group is displaced and thus prevents or at leastdelays the hydrolysis of the phospholipid.

Hence in a preferred manner according to the invention the matrix, inaddition to PS and PC, contains further at least one fat component andeven more preferably a fat component and a wax component and in anotherembodiment preferably at least one polyalcohol component.

According to the invention particularly suitable matrix materials aresubstances which enable a complete encapsulation to develop as well assubstances which provide a matrix of high stability and low shearstress.

In a preferred embodiment the claimed matrix has a water-containing coatwhich can also be permanently flexible.

In this context the coat which is preferably also composed of gelatin,glycerol, sugar(alcohols), starch, polysaccharides and mixtures thereofshould have a water content of 1.0 to 10.0% by weight based on the totalcoat. Sorbitol is particularly preferred as a sugar alcohol for the coatand carragenans, alginates and/or pectins are particularly preferred aspolysaccharide components. Finally the invention also provides that thecoat of the matrix contains silicon dioxide, calcium carbonate, dyesthat are suitable for foods, colour pigments and/or talcum as furtheradditives.

Depending on the composition of the bioactive component and the class ofsubstances that make up the coating material, the weight ratio of coatand the bioactive component can be important for the product quality ofthe total matrix according to the invention. In this context weightratios of the coat to bioactive component have proven to be particularsuitable which are between 1:0.25 to 10.0 and particularly preferably1:1 to 5.0.

The total diameter of the matrix which should be in particular between0.3 and 20 mm according to the present invention also depends on therespective intended use.

In addition to the matrix itself the present invention also encompassesits use whereby on the basis of the component contained in the matrixespecially strength being the ability to cope with mental or/andphysical stress and functional capacity, the prevention of elevatedlevels of serum cholesterol, the promotion or/and preservation of healthand quite generally the improvement of well-being are of primaryimportance.

Hence with the matrix according to the invention a formulation ofphosphatidyl serine and phosphatidyl choline has been found which can bemanufactured in an economically acceptable manner and which, in contrastto the other known formulations, has a pronounced oxidative andhydrolytic stability. Thus the matrix according to the inventioncombines two features which were previously not compatible, namely theability to be encapsulated and at the same time to have an adequatestability. This combination is especially pronounced in the case ofmatrices with thixotropic or shear diluting properties. However, it alsooccurs in matrix forms which are pasty and behave in a Newtonian manner.

The following examples elucidate these advantages of the matrix with abioactive component containing phospholipid according to the invention.

EXAMPLE Example 1

The hard matrix contained 20% by weight phosphatidyl serine and 15% byweight phosphatidyl choline as the bioactive component. Other componentswere each 3% by weight phosphatidyl inositol and 2% by weightphosphatidyl ethanolamine, 38% by weight of a mixture of refined soya,two partially hydrogenated soya oils of different melting points as wellas 3% by weight of a bee wax, 2% by weight of a mixture composed ofvitamins E and D, tocotrieols and β-carotin. The remainder making up100% by weight was composed of the typical substances accompanyinglecithins such as glycolipids, phytosterols and oligosugars.

The two bioactive components were homogenously dispersed in the matrixwhich was present as pellets having a diameter of 3 to 8 mm.

Example 2

The hard matrix contained 20% by weight phosphatidyl serine and 5% byweight phosphatidyl choline and 4% by weight phosphatidyl inositol asthe bioactive component. Other components were 6% by weight phosphatidylethanolamine and 2% by weight phosphatidic acid, 45% by weight palmkernel oil and 5% by weight of a bee wax as well as 0.2% by weightvitamin E. The remainder making up 100% by weight was composed of thetypical substances accompanying lecithins such as glycolipids,phytosterols and oligosugars.

Example 3

The hard matrix contained 30% by weight phosphatidyl serine, 4% byweight phosphatidyl choline and 2% by weight phosphatidyl inositol asthe bioactive component. Other components were each 4% by weightphosphatidyl ethanolamine and 1% by weight phosphatidic acid, 45% byweight palm kernel oil and 5% by weight of a bee wax as well as 0.2% byweight vitamin E. The remainder making up 100% by weight was composed ofthe typical substances accompanying lecithins such as glycolipids,phytosterols and oligosugars.

Stability of the Phospholipids:

Table 1 shows that in the case of phosphatidyl serine (PS) that is verysensitive to hydrolysis and was encapsulated in a soft gelatin capsuleas an example, embedding the phospholipids in the matrix according tothe invention has a stabilizing effect towards hydrolysis among others.

Two phospholipid-containing lecithins in a standard preparationcorresponding to the prior art served as a comparison which wereoptimized with regard to their ability to be encapsulated by thestandard rotary die process. The lecithins were stored in anencapsulated form at room temperature (23° C.).

Examples 1 to 3 are the three described inventive examples; examples 4to 6 are comparative examples. TABLE 1 Ex- initial value after 3 monthsafter 6 months after 12 months amples PS [%] [%] [%] [%] 1 100 99 98 972 100 99 97 96 3 100 99 98 96 4 100 84 80 69 5 100 93 79 70 6 100 75 6462

1. A matrix with a bioactive component containing phospholipid, thematrix containing 5 to 98% by weight phosphatidyl serine (PS) and 1 to90% by weight phosphatidyl choline (PC) as the bioactive component andin addition 1 to 94% by weight of at least one other component from theseries comprising fat component of vegetable and/or animal origin, waxcomponent, polyalcohol component and other physiologically compatibleadditives, the other matrix components being selected such that thetotal matrix is solid or paste-like at room temperature and exhibitsproperty of shear dilution.
 2. The matrix as claimed in claim 1, whereinthe bioactive component contains 10 to 40% by weight of phosphatidylserine.
 3. The matrix as claimed in claim 1, wherein the bioactivecomponent contains 2.0 to 20% by weight of phosphatidyl choline.
 4. Thematrix as claimed in claim 1, containing 10 to 70% by weight of the fatcomponent and/or 3 to 30% by weight of the wax component and/or 1 to 30%by weight of polyalcohol component and/or 1 to 5% by weight of otherphysiologically compatible additives.
 5. The matrix as claimed in claim1, wherein it contains refined, hydrogenated and/or fractionated fatsand in particular those that are rich in omega-3 and/or omega-6 fattyacids such as docosahexaenoic acid, arachidonic acid, eicosapentaenoicacid and conjugated linolenic acid, free fatty acids, in particularomega-3 and omega-6 fatty acids, bee wax, candellila wax, shellac,paraffin, monoglycerides or diglycerides, polyethylene glycol,polysorbate, polyglycerol esters, sugar esters or sorbitan esters,tocopherols and derivatives thereof, tocotrienols and derivativesthereof, polycosanols and derivatives thereof, vitamins such as vitaminC and E also in a derivatized form, amino acids in particular theessential, branched and non-proteinogenic amino acids such as theanine,amino acid derivatives such as creatine, taurine, carnitine,phytosterols and derivatives thereof, (poly)phenolic compounds andderivatives thereof such as catechol, phenolic acids such as gallicacid, hydroxycinnamic acids, coumarins, (iso)flavonoids such asquercetin or genistein, lignans and lignins as well as tannin, saponins,mono-, sesqui- and di-terpenes, carotinoids such as beta-carotin, luteinor lycopin, glucosinolates, roughage such as non-starch polysaccharides,extracts of vegetable and/or animal origin, physiologically activeproteins such as lactoferrin and glycomacropeptide, glycolipids such assphingosine or (phyto)sphingomyelin and/or mineral components.
 6. Thematrix as claimed in claim 1, wherein the matrix has a water-containingcoat.
 7. The matrix as claimed in claim 6, wherein the coat has a watercontent of 1.0 to 10.0% by weight based on total coat.
 8. The matrix asclaimed in claim 6, wherein the coat is composed of gelatin, glycerol,sugar (alcohols), starch, polysaccharides and mixtures thereof.
 9. Thematrix as claimed in claim 8, wherein the coat contains sorbitol assugar alcohol and carrageenans, alginates and/or pectins aspolysaccharide.
 10. The matrix as claimed in claim 8, wherein the coatcontains silicon dioxide, calcium carbonate, dyes that are suitable forfoods, colour pigments and/or talcum as further additives.
 11. Thematrix as claimed in claim 6, wherein the weight ratio of the coat tobioactive component is 1:0.25 to 10.0.
 12. The matrix as claimed inclaim 1, having a total diameter of 0.3 to 20 mm.
 13. (canceled) 14.(canceled)
 15. Method for strengthening ability to cop with mentaland/or physical stress and functional capacity, for improvingwell-being, for promoting and/or preserving health and for preventingelevated levels of serum cholesterol in a subject, comprising steps of:a) providing a matrix as claimed in claim 1; b) producing apharmaceutical preparation comprising said matrix; and c) administeringan effective amount of said pharmaceutical preparation to said subject.16. The matrix as claimed in claim 1, wherein the bioactive componentcontains 15 to 30% by weight of phosphatidyl serine.
 17. The matrix asclaimed in claim 1, containing 20 to 50% by weight of the fat componentand/or 5 to 20% by weight of the wax component and/or 2 to 20 % byweight of polyalcohol component and/or 1 to 5% by weight of otherphysiologically compatible additives.
 18. The matrix as claimed in claim6, wherein the weight ratio of the coat to bioactive component is 1:1 to5.0.